TREATMENT WITH H2O2, RESVERATROL, OR NICOTINAMIDE

To evaluate the effects of H₂O₂ on cell viability of BM-MSCs, cells were exposed to different dosages of H₂O₂ ranging from 25 to 400 μM for 72 h. At the specific time points, BM-MSCs were analyzed by both qualitative and quantitative cell viability assays. To evaluate the influence of H₂O₂ on the stages of osteogenic differentiation, BM-MSCs were treated with 25 μM, 50 μM, and 100 μM H₂O₂ for 21 days. Untreated cells served as a control. To investigate the role of SIRT1 in the early stage of osteogenesis, BM-MSCs were treated with 5 μM ResV in the presence of 100 μM H₂O₂ for 7 days and subjected to subsequent experiments. To investigate the role of SIRT1 in the late stage of osteogenesis, BM-MSCs were treated with 10 mM NAM in the presence of 50 μM H₂O₂ for the last 7 days and subjected to subsequent experiments.